Application of loop-mediated isothermal amplification (lamp) in microbiological water quality analysis
Abstract
Water is life thus access to safe drinking water is essential to health and a basic human right.
Kenya is considered a water scarce country whose water sources are becoming more
contaminated through changes in land use and poor solid waste management. This has in turn
affected the quantity and quality of portable water available. There are nearly 1.7 billion cases of
waterborne diarrheal disease every year with over 760 000 deaths of children under five years.
Traditional methods of identification of waterborne pathogens are less specific, sensitive, timeconsuming
and laborious, so there is a need for the development of innovative methods for their
rapid identification. Recent advances in molecular cloning and recombinant DNA techniques
have revolutionized the detection of pathogens in foods. In this study the application of a LAMP
based technique for the rapid identification of the Escherichia coli, Enterococcus feacalis and
Clostridia perfringens DNAs was undertaken. Suitable primers were designed based on specific
gene MalB of Escherichia coli, tuf gene of Enterococcus feacalis and CPE gene of Clostridia
perfringens for amplification. Selective media for the various indicator microbes were used and
specific primers for both LAMP and PCR designed. The study reveals that bottled water from
Majesty and Starpop companies were contaminated by coliforms while the ground water from
Kikuyu Springs, rainwater from Lang’ata and tapped water from Naivasha’s Fishermans
campsite were contaminated by the three target bacteria. This was possible via culture work as
LAMP and PCR assays were not fully optimized to enable the detection of Escherichia coli,
Enterococcus feacalis and Clostridiun perfrigens DNAs. The study shows LAMP as a viable tool
in microbial water quality analysis from the little amplification achieved with the positive
controls that had high DNA concentrations
Citation
Master of Science in Biotechnology, University of Nairobi, 2014Publisher
University of Nairobi